Information on EC 2.7.7.52 -

Information on EC 2.7.7.52 - RNA uridylyltransferase:

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EC NUMBER	COMMENTARY
2.7.7.52	-

RECOMMENDED NAME	GeneOntology No.
RNA uridylyltransferase	GO:0050265

REACTION	REACTION DIAGRAM	COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
UTP + RNAn = diphosphate + RNAn+1		enzyme requires an oligoribonucleotide or polyribonucleotide with a free terminal 3'-OH as a primer	-	-	-
UTP + RNAn = diphosphate + RNAn+1		mechanism, uridine insertion in the editing process	Trypanosoma brucei	-	643506

REACTION TYPE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
nucleotidyl group transfer	-	-	-	-

PATHWAY	KEGG Link	MetaCyc Link
No entries in this field

SYSTEMATIC NAME	IUBMB Comments
UTP:RNA uridylyltransferase	The enzyme requires an oligoribonucleotide or polyribonucleotide with a free terminal 3'-OH as a primer.

SYNONYMS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
3' terminal uridylyl transferase	Trypanosoma brucei	-	-	643503, 643505
RET	Trypanosoma brucei	-	i.e. RNA editing TUTase	643505, 643506
RET1	Leishmania tarentolae, Trypanosoma brucei	-	-	675809, 691064
RET2	Trypanosoma brucei	-	-	675809, 691064
TbMP57 TUTase	Trypanosoma brucei	-	-	677097
TbTUT3	Trypanosoma brucei	-	-	661742
TbTUT4	Trypanosoma brucei	Q381M1	-	675434
terminal RNA uridylyltransferase	Trypanosoma brucei	-	-	675809, 691064
terminal RNA uridylyltransferase	Leishmania tarentolae	-	-	675809
terminal U transferase (TUTase)	Trypanosoma brucei	-	-	677097
terminal uridylyltransferase	-	-	-	-
terminal uridylyltransferase	Trypanosoma brucei	Q381M1	-	675434
TUT	-	-	-	-
TUT1	Leishmania tarentolae	-	-	691064
TUT4	Trypanosoma brucei	Q381M1	-	675434
TUTase	Trypanosoma brucei	-	-	675434, 675809, 691064
TUTase	Leishmania tarentolae	-	-	675809, 691064
TUTase 1	Trypanosoma brucei	-	-	662323, 691064
TUTase 2	Trypanosoma brucei	-	-	691064
TUTase 3	Trypanosoma brucei	-	-	691064
TUTase 4	Trypanosoma brucei	-	-	691064
U6-TUTase	Homo sapiens	-	U6 snRNA-specific terminal uridylyltransferase	643504
uridylyltransferase, terminal	-	-	-	-
3'-terminal uridylyl transferase 1	Trypanosoma brucei	-	-	662323
additional information	Leishmania tarentolae, Trypanosoma brucei	-	enzyme is a member of the DNA polymerase beta nucleotidyltransferase superfamily	643502
additional information	Trypanosoma brucei	-	the enzyme belongs to a large enzyme superfamily typified by DNA polymerase beta	675434, 691064
additional information	Leishmania tarentolae	-	the enzyme belongs to a large enzyme superfamily typified by DNA polymerase beta	691064

CAS REGISTRY NUMBER	COMMENTARY
78519-53-6	-

ORGANISM	COMMENTARY	LITERATURE	SEQUENCE CODE	SEQUENCE DB	SOURCE
Crithidia fasciculata	-	643501	-	-
Homo sapiens	-	643500	-	-
Homo sapiens	an unspecific enzyme form and an enzyme form specific for U6 sn-RNA	643504	-	-
Leishmania tarentolae	-	643502, 675809, 691064	-	-
Leishmania tarentolae	UC strain	1997	-	-
Leishmania tarentolae UC	UC strain	1997	-	-
Trypanosoma brucei	-	643502, 661742, 662323, 675809, 691064	-	-
Trypanosoma brucei	; isozyme TUT4	675434	Q381M1	SwissProt
Trypanosoma brucei	editosome protein TbMP57, distinct from mitochondrial enzyme form, i.e. TbTUTase 108	643503	-	-
Trypanosoma brucei	RET1 and RET2, i.e. RNA editing TUTases 1 and 2, as components of different editing complexes	643505, 643506	-	-
Trypanosoma brucei	strain TREU-667	677097	-	-
Trypanosoma brucei TREU-667	strain TREU-667	677097	-	-
Vigna unguiculata	-	643498	-	-

GENERAL INFORMATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

SUBSTRATE	PRODUCT	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY/ Substrate	LITERATURE/ Substrate	COMMENTARY/ Product	LITERATURE/ Product	Reversibility r=reversible ir=irreversible ?=not specified
ATP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	low activity	643502	-	-	?
ATP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	editosomal enzyme form, no activity	643503	-	-	-
CTP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	-	643502	-	-	?
CTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	editosomal enzyme form, no activity	643503	-	-	-
GTP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	about 50% of the activity with UTP, prefers Mn2+ as divalent cation	643502	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	-	643502	-	643502	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	-	661742, 675809	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	-	1997	-	1997	?
UTP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	-	675809	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	Q381M1	-	675434	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Vigna unguiculata	-	the enzyme requires a single-stranded oligoribonucleotide or polyribonucleotide with a free terminal 3'-OH as primer, e.g. oligoA20, tRNAAsp, E. coli RNA, alfalafa mosaic virus RNA 4	643498	-	643498	?
UTP + RNAn	diphosphate + RNAn+1		Crithidia fasciculata	-	small guide RNA, i.e. gRNA, marked specificity for UTP	643501	-	643501	?
UTP + RNAn	diphosphate + RNAn+1		Vigna unguiculata	-	marked specificity for UTP	643498	-	643498	?
UTP + RNAn	diphosphate + RNAn+1		Homo sapiens	-	3'-poly(A) of virion RNA	643500	-	643500	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	editosomal enzyme form shows preference for a 3' terminal A or G, while the mitochondrial enzyme form does not, mitochondrial enzyme form adds Us at the 3' and the 5' end of the RNA	643503	-	643503	?
UTP + RNAn	diphosphate + RNAn+1		Leishmania tarentolae	-	RNA substrate specificity, overview	643502	-	643502	?
UTP + RNAn	diphosphate + RNAn+1		Vigna unguiculata	-	RNA substrate specificity, overview	643498	-	643498	?
UTP + RNAn	diphosphate + RNAn+1		Homo sapiens	-	specifically modifies the 3'-UMP terminal of mammalian U6 small nuclear RNA, i.e. snRNA, structural requirements and specificity, overview	643504	-	643504	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	kinetoplast RNA	643506	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	the mitochondrial enzyme adds a single U to the 3'-end of single-stranded RNA	643505	-	643505	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	involved in uridine insertion in the editing process of RNA	643506	-	643506	?
UTP + RNAn	diphosphate + RNAn+1		Vigna unguiculata	-	RNA uridylyltransferase might function in uridylating specific proteins, RNA is not a natural substrate	643498	-	643498	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	responsible for post-transcriptional RNA editing process of mitochondrial transcripts in kinetoplastid protozoans	643506	-	643506	?
UTP + RNAn	diphosphate + RNAn+1		Crithidia fasciculata	-	responsible for post-transcriptional RNA editing process of mitochondrial transcripts in kinetoplastid protozoans	643501	-	643501	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of non-coded poly(U) tail to gRNAs	643505	-	643505	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of non-coded poly(U) tail to gRNAs	643506	-	643506	?
UTP + RNAn	diphosphate + RNAn+1		Crithidia fasciculata	-	addition of non-coded poly(U) tail to gRNAs	643501	-	643501	?
UTP + RNAn	diphosphate + RNAn+1		Homo sapiens	-	acts as a host factor to initiate RNA synthesis by poliovirus RNA polymerase in vitro	643500	-	643500	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of primarily single U to single-stranded RNA, addition of the number of Us specified by a guide RNA to insertion editing-like substrates	643503	-	643503	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	TbMP57 TUTase, an enzyme thought to act exclusively in U-insertion, can also function within the U-deletion cycle, after cleavage and 3'-U-exo but before U-deletional cycle	677097	-	-	?
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	Q381M1	a template-independent RNA nucleotidyltransferases that specifically recognize UTP, it possesses conserved catalytic and UTP recognition domains. A single nucleoside triphosphate is bound in the active site by a complex network of interactions between amino acid residues, a magnesium ion and highly ordered water molecules with the UTP�s base, ribose and phosphate moieties, structure-function analysis, overview	675434	-	-	?
GTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	editosomal enzyme form, no activity	643503	-	-	-
additional information	?	-	Trypanosoma brucei	-	-	643502	-	-	-
additional information	?	-	Trypanosoma brucei	-	reduction of isozyme RET1 leads to decrease in edited RNA and inhibited growth, lowers the uridine insertion and leads predominantly to shorter gRNAs, down-regulation of isozyme RET2 inhibits growth and in vivo uridine insertion, but has no effect on the length of gRNAs	643506	-	-	-
additional information	?	-	Leishmania tarentolae	-	enzyme interacts with a minor fraction of total RNA ligase	643502	-	-	-
additional information	?	-	Trypanosoma brucei	-	many editing changes are developmentally regulated	643506	-	-	-
additional information	?	-	Trypanosoma brucei	-	down-regulation of RET1, but not of RET2, affects length distribution of gRNA 3' oligo(U) tails	643505	-	-	-
additional information	?	-	Trypanosoma brucei	-	Inhibition of RNA editing by down-regulation of expression of the mitochondrial RNA editing TUTase 1 by RNA interference has profound effects on kinetoplast biogenesis in Trypanosoma brucei procyclic cells. De novo synthesis of the apocytochrome b and cytochrome oxidase subunit I proteins is no longer detectable after 3 days of RNAi. The effect on protein synthesis correlates with a decline in the levels of the assembled mitochondrial respiratory complexes III and IV, and also with cyanide-sensitive oxygen uptake. The steady-state levels of nuclear-encoded subunits of complexes III and IV are also significantly decreased. Because the levels of the corresponding mRNAs are not affected, the observed effect is likely due to an increased turnover of these imported mitochondrial proteins. This induced protein degradation is selective for components of complexes III and IV, because little effect is observed on components of the F1-F0 -ATPase complex and on several other mitochondrial proteins	662323	-	-	-
additional information	?	-	Trypanosoma brucei, Leishmania tarentolae	-	post-transcriptional uridylylation of guide RNAs by RNA editing TUTase 1 or RET1, a multi-functional RNA processing enzyme, and U-insertion mRNA editing by RNA editing TUTase 2 or RET2, biological functions of TUT isozymes, RNA processing in mitochondria of trypanosomes, detailed overview	691064	-	-	-
additional information	?	-	Trypanosoma brucei	Q381M1	TUT4 substrate specificity toward nucleoside triphosphate substrates with a synthetic 5'-radiolabeled 24-mer RNA as a primer, in presence of Mg2+ ions TUT4 preferentially incorporates uridylyl residues but the reaction with CTP is also detectable, TbTUT4 incorporates only one deoxynucleotide into RNA, overview. UTP binding site structure and structure-activity analysis	675434	-	-	-
additional information	?	-	Leishmania tarentolae	-	UTP recognition mechanism, overview. The NTD-CTD bi-domain catalytic modules shared by TUTases and non-canonical poly(A) polymerases, ncPAPs, are quite promiscuous in NTP binding, NTP selectivity of TUTase-like catalytic modules, RNA specificity, overview	691064	-	-	-
additional information	?	-	Trypanosoma brucei	-	UTP recognition mechanism, overview. UTP specificity is determined primarily by the two closely positioned carboxylic residues, D297/D421 and E300/E424, which coordinate a crucial water molecule indicated in TUT4/RET2, uracil base interactions with invariant amino acids N147, S148, Y189, D297, E300 of TUT4. The NTD-CTD bi-domain catalytic modules shared by TUTases and non-canonical poly(A) polymerases, ncPAPs, are quite promiscuous in NTP binding, NTP selectivity of TUTase-like catalytic modules, RNA specificity, overview	691064	-	-	-

NATURAL SUBSTRATES	NATURAL PRODUCTS	REACTION DIAGRAM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY SUBSTRATE	LITERATURE (Substrate)	COMMENTARY PRODUCT	LITERATURE (Product)
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	-	643502	-	643502
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	Q381M1	-	675434	-	-
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	involved in uridine insertion in the editing process of RNA	643506	-	643506
UTP + RNAn	diphosphate + RNAn+1		Vigna unguiculata	-	RNA uridylyltransferase might function in uridylating specific proteins, RNA is not a natural substrate	643498	-	643498
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	responsible for post-transcriptional RNA editing process of mitochondrial transcripts in kinetoplastid protozoans	643506	-	643506
UTP + RNAn	diphosphate + RNAn+1		Crithidia fasciculata	-	responsible for post-transcriptional RNA editing process of mitochondrial transcripts in kinetoplastid protozoans	643501	-	643501
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of non-coded poly(U) tail to gRNAs	643505	-	643505
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of non-coded poly(U) tail to gRNAs	643506	-	643506
UTP + RNAn	diphosphate + RNAn+1		Crithidia fasciculata	-	addition of non-coded poly(U) tail to gRNAs	643501	-	643501
UTP + RNAn	diphosphate + RNAn+1		Homo sapiens	-	acts as a host factor to initiate RNA synthesis by poliovirus RNA polymerase in vitro	643500	-	643500
UTP + RNAn	diphosphate + RNAn+1		Trypanosoma brucei	-	addition of primarily single U to single-stranded RNA, addition of the number of Us specified by a guide RNA to insertion editing-like substrates	643503	-	643503
additional information	?	-	Trypanosoma brucei	-	-	643502	-	-
additional information	?	-	Trypanosoma brucei	-	reduction of isozyme RET1 leads to decrease in edited RNA and inhibited growth, lowers the uridine insertion and leads predominantly to shorter gRNAs, down-regulation of isozyme RET2 inhibits growth and in vivo uridine insertion, but has no effect on the length of gRNAs	643506	-	-
additional information	?	-	Leishmania tarentolae	-	enzyme interacts with a minor fraction of total RNA ligase	643502	-	-
additional information	?	-	Trypanosoma brucei	-	many editing changes are developmentally regulated	643506	-	-
additional information	?	-	Trypanosoma brucei	-	down-regulation of RET1, but not of RET2, affects length distribution of gRNA 3' oligo(U) tails	643505	-	-
additional information	?	-	Trypanosoma brucei	-	Inhibition of RNA editing by down-regulation of expression of the mitochondrial RNA editing TUTase 1 by RNA interference has profound effects on kinetoplast biogenesis in Trypanosoma brucei procyclic cells. De novo synthesis of the apocytochrome b and cytochrome oxidase subunit I proteins is no longer detectable after 3 days of RNAi. The effect on protein synthesis correlates with a decline in the levels of the assembled mitochondrial respiratory complexes III and IV, and also with cyanide-sensitive oxygen uptake. The steady-state levels of nuclear-encoded subunits of complexes III and IV are also significantly decreased. Because the levels of the corresponding mRNAs are not affected, the observed effect is likely due to an increased turnover of these imported mitochondrial proteins. This induced protein degradation is selective for components of complexes III and IV, because little effect is observed on components of the F1-F0 -ATPase complex and on several other mitochondrial proteins	662323	-	-
additional information	?	-	Trypanosoma brucei, Leishmania tarentolae	-	post-transcriptional uridylylation of guide RNAs by RNA editing TUTase 1 or RET1, a multi-functional RNA processing enzyme, and U-insertion mRNA editing by RNA editing TUTase 2 or RET2, biological functions of TUT isozymes, RNA processing in mitochondria of trypanosomes, detailed overview	691064	-	-

COFACTOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

METALS and IONS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
Ca2+	Trypanosoma brucei	Q381M1	influences the TUT4 conformation	675434
K+	Trypanosoma brucei	Q381M1	-	675434
KCl	Trypanosoma brucei	-	stimulated by concentrations up to 50 mM	661742
Mg2+	Vigna unguiculata	-	divalent cation required, Mg2+ or Mn2+, 4-12 mM stimulate, optimal at 5 mM	643498
Mg2+	Crithidia fasciculata	-	required	643501
Mg2+	Leishmania tarentolae	-	required	643502
Mg2+	Homo sapiens	-	dependent on	643504
Mg2+	Trypanosoma brucei	-	optimum: 0.5 mM	661742
Mg2+	Trypanosoma brucei	Q381M1	increases the UTP substrate specificity of TUT4 and influences the enzyme conformation; maximal activity at 2 mM Mg2+	675434
Mg2+	Leishmania tarentolae, Trypanosoma brucei	-	assay with 10 mM Mg2+	675809
Mn2+	Vigna unguiculata	-	divalent cation required, Mg2+ or Mn2+, 1.1-1.5 mM stimulate	643498
Mn2+	Leishmania tarentolae	-	requires Mn2+ or Mg2+; selectively inhibits the distributive activity of the enzyme; when GTP, CTP, or ATP are used as substrates, Mn2+ is more effective than Mg2+	643502
Mn2+	Trypanosoma brucei	Q381M1	replacement of magnesium with manganese leads to a dramatic stimulation of processivity and partial loss of selectivity for UTP	675434
additional information	Trypanosoma brucei	Q381M1	divalent cations essential for catalysis	675434

INHIBITORS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
Aurintricarboxylic acid	Vigna unguiculata	-	-	643498	2D-image
brRNA	Leishmania tarentolae	-	inhibits the number of added residues	643502	-
Cibacron blue F3GA	Vigna unguiculata	-	-	643498	2D-image
diphosphate	Vigna unguiculata	-	-	643498	2D-image
heparin	Leishmania tarentolae	-	complete inhibition of solubilized enzyme	1997	2D-image
heparin	Vigna unguiculata	-	-	643498	2D-image
Ionic strength	Vigna unguiculata	-	quite sensitive to ionic strength, activity decreases by 50% at about 40 mM (NH4)2SO4 or 125 mM potassium acetate	643498	-
additional information	Vigna unguiculata	-	not inhibitory: actinomycin C, rifamycin, alpha-amanitin, phosphate	643498	-
additional information	Trypanosoma brucei	-	RNA interference, i.e. RNAi, induced by tetracyclines causes down-regulation of RET1 and RET2 expression in vitro and in vivo	643502, 643505	-

ACTIVATING COMPOUND	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
Ionic strength	Homo sapiens	-	stimulates, e.g. 600 mM KCl	643504	-
protein TbMP81	Trypanosoma brucei	-	editosome protein, interaction enhances the enzyme activity	643503	-

KM VALUE [mM]	KM VALUE [mM] Maximum	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
0.0002	-	RNA	Trypanosoma brucei	Q381M1	; mutant S148A	675434	2D-image
0.0003	-	RNA	Trypanosoma brucei	Q381M1	mutant S188A	675434	2D-image
0.01	-	RNA	Trypanosoma brucei	Q381M1	mutant R307A	675434	2D-image
0.0726	-	RNA	Trypanosoma brucei	Q381M1	mutant R141A	675434	2D-image
0.132	-	RNA	Trypanosoma brucei	Q381M1	mutant R121A	675434	2D-image
0.0004	-	UTP	Trypanosoma brucei	Q381M1	mutant R307A	675434	2D-image
0.001	-	UTP	Trypanosoma brucei	Q381M1	; mutant R141A	675434	2D-image
0.0028	-	UTP	Trypanosoma brucei	Q381M1	mutant R121A	675434	2D-image
0.0139	-	UTP	Trypanosoma brucei	Q381M1	mutant D297N	675434	2D-image
0.0213	-	UTP	Trypanosoma brucei	Q381M1	mutant Y189F	675434	2D-image
0.034	-	UTP	Trypanosoma brucei	Q381M1	mutant S188A	675434	2D-image
0.0873	-	UTP	Trypanosoma brucei	Q381M1	mutant S148A	675434	2D-image
0.1153	-	UTP	Trypanosoma brucei	Q381M1	mutant N147A	675434	2D-image
0.1728	-	UTP	Trypanosoma brucei	Q381M1	mutant D297A	675434	2D-image
0.3084	-	UTP	Trypanosoma brucei	Q381M1	mutant E300A	675434	2D-image
36	48	UTP	Trypanosoma brucei	-	-	661742	2D-image
additional information	-	additional information	Trypanosoma brucei	Q381M1	steady-state enzyme kinetics, overview	675434	-

TURNOVER NUMBER [1/s]	TURNOVER NUMBER MAXIMUM[1/s]	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
0.00166	-	RNA	Trypanosoma brucei	Q381M1	mutant S188A	675434	2D-image
0.005	-	RNA	Trypanosoma brucei	Q381M1	mutant R121A	675434	2D-image
0.00833	-	RNA	Trypanosoma brucei	Q381M1	mutant R307A	675434	2D-image
0.0166	-	RNA	Trypanosoma brucei	Q381M1	mutant S148A	675434	2D-image
0.0216	-	RNA	Trypanosoma brucei	Q381M1	mutant R141A	675434	2D-image
3.3e-05	-	UTP	Trypanosoma brucei	Q381M1	mutant R121A	675434	2D-image
6.66e-05	-	UTP	Trypanosoma brucei	Q381M1	mutant D297N	675434	2D-image
0.00011	-	UTP	Trypanosoma brucei	Q381M1	mutant R141A	675434	2D-image
0.000133	-	UTP	Trypanosoma brucei	Q381M1	mutant R307A	675434	2D-image
0.00033	-	UTP	Trypanosoma brucei	Q381M1	mutant D297A	675434	2D-image
0.0005	-	UTP	Trypanosoma brucei	Q381M1	mutant S188A	675434	2D-image
0.005	-	UTP	Trypanosoma brucei	Q381M1	mutant N147A	675434	2D-image
0.0066	-	UTP	Trypanosoma brucei	Q381M1	mutant E300A	675434	2D-image
0.015	-	UTP	Trypanosoma brucei	Q381M1	mutant Y189F	675434	2D-image
0.021	-	UTP	Trypanosoma brucei	Q381M1	mutant S148A	675434	2D-image

kcat/KM VALUE [1/mMs^-1]	kcat/KM VALUE [1/mMs^-1] Maximum	SUBSTRATE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

Ki VALUE [mM]	Ki VALUE [mM] Maximum	INHIBITOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

IC50 VALUE [mM]	IC50 VALUE [mM] Maximum	INHIBITOR	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	IMAGE
No entries in this field

SPECIFIC ACTIVITY [µmol/min/mg]	SPECIFIC ACTIVITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
6.67	-	Leishmania tarentolae	-	purified enzyme	643502
8.2	-	Leishmania tarentolae	-	purified recombinant enzyme	643502
additional information	-	Vigna unguiculata	-	-	643498
additional information	-	Homo sapiens	-	-	643500

pH OPTIMUM	pH MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
6.8	-	Leishmania tarentolae	-	assay at	1997
7.9	-	Crithidia fasciculata	-	assay at	643501
7.9	-	Homo sapiens	-	assay at	643504
8	-	Trypanosoma brucei	Q381M1	assay at	675434
8	-	Leishmania tarentolae, Trypanosoma brucei	-	assay at	675809
8.2	-	Vigna unguiculata	-	assay at	643498

pH RANGE	pH RANGE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

TEMPERATURE OPTIMUM	TEMPERATURE OPTIMUM MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
27	-	Leishmania tarentolae	-	assay at	1997
27	-	Trypanosoma brucei	Q381M1	assay at	675434
27	-	Leishmania tarentolae, Trypanosoma brucei	-	assay at	675809
30	-	Vigna unguiculata	-	-	643498
30	-	Homo sapiens	-	assay at	643504

TEMPERATURE RANGE	TEMPERATURE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

pI VALUE	pI VALUE MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

SOURCE TISSUE	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE	SOURCE
HeLa cell	Homo sapiens	-	-	643500, 643504
leaf	Vigna unguiculata	-	-	643498
procyclic form	Trypanosoma brucei	-	-	662323
trypanosome	Leishmania tarentolae, Trypanosoma brucei	-	-	691064

LOCALIZATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	GeneOntology No.	LITERATURE	SOURCE
cytosol	Trypanosoma brucei	-	TUT3 and TUT4	5829	691064
kinetoplast	Crithidia fasciculata	-	-	20023	643501
kinetoplast	Trypanosoma brucei	-	-	20023	643506
membrane	Vigna unguiculata	-	-	16020	643498
mitochondrion	Leishmania tarentolae	-	-	5739	1997, 643502, 675809
mitochondrion	Crithidia fasciculata	-	-	5739	643501
mitochondrion	Trypanosoma brucei	-	-	5739	643502, 643503
mitochondrion	Trypanosoma brucei	-	2 isozymes RET1 and RET2 in mitochondria editosomes	5739	643506
mitochondrion	Leishmania tarentolae	-	RET1	5739	691064
mitochondrion	Trypanosoma brucei	-	TUT1 and TUT2	5739	691064
additional information	Trypanosoma brucei	-	integral editosome protein, distinct from mitochondrial enzyme form	-	643503
additional information	Trypanosoma brucei	-	isozymes are located in 2 different complexes	-	643506
additional information	Trypanosoma brucei	-	non-mitochondrial localization	-	661742

PDB	SCOP	CATH	ORGANISM
2e5g, download	SCOP (2e5g)	CATH (2e5g)	Homo sapiens
2ikf, download	SCOP (2ikf)	CATH (2ikf)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2nom, download	SCOP (2nom)	CATH (2nom)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2q0c, download	SCOP (2q0c)	CATH (2q0c)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2q0d, download	SCOP (2q0d)	CATH (2q0d)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2q0e, download	SCOP (2q0e)	CATH (2q0e)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2q0f, download	SCOP (2q0f)	CATH (2q0f)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)
2q0g, download	SCOP (2q0g)	CATH (2q0g)	Trypanosoma brucei brucei (strain 927/4 GUTat10.1)

MOLECULAR WEIGHT	MOLECULAR WEIGHT MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
37800	-	Trypanosoma brucei	Q381M1	gel filtration	675434
43000	-	Crithidia fasciculata	-	gel filtration	643501
50500	-	Vigna unguiculata	-	velocity sedimentation	643498
68000	-	Homo sapiens	-	gel filtration	643500
135000	-	Leishmania tarentolae	-	native PAGE	643502
500000	-	Trypanosoma brucei	-	native PAGE and gel filtration	643502
500000	-	Trypanosoma brucei	-	mitochondrial enzyme form, gel filtration	643503
700000	-	Trypanosoma brucei	-	RNA-stabilized higher complexed form, native PAGE and glycerol gradient sedimentation	643502

SUBUNITS	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
?	Homo sapiens	-	x * 115000, U6-TUTase, SDS-PAGE	643504
?	Trypanosoma brucei	Q381M1	x * 37800, TUT4, crystal structure analysis	675434
tetramer	Leishmania tarentolae	-	4 * 140000, TUT1	691064
homotetramer	Leishmania tarentolae	-	-	675809
additional information	Trypanosoma brucei	Q381M1	TbTUT4 represents a minimal catalytically active RNA uridylyltransferase consisting of only two domains that define the catalytic center at the bottom of the nucleoside triphosphate and RNA substrate binding cleft. The enzyme shows two significantly different conformations: one molecule is in a relatively open apo conformation, whereas the other displays a more compact TUTase-UTP complex, structure-function analysis, overview	675434
additional information	Trypanosoma brucei	-	in RET1, an extra 150 amino acid insertion constitutes a helix-rich domain essential for activity, the insertion is designated as middle domain, occurs at a conserved site within the catalytic domain and is found in RET1, RET2 and TUT3, but not in TUT4, analysis of Structural diversity of trypanosomal TUTases, overview	691064

POSTTRANSLATIONAL MODIFICATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

Crystallization/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
purified recombinant N-terminally His-tagged TUT4, free or with bound 2'-deoxyribonucleoside, at a concentration of 10 mg/mL in 10 mM HEPES, pH 7.6, 70 mM KCl, 0.5 mM DTT, and in presence of 4 mM MgCl2 and 0.05 mM UTP, screening with commercial crystallization screens, vapor diffusion technique, 4�C, cryoprotection by 15% glycerol, X-ray diffraction structure determination and analysis at 2.0-2.4 A resolution; small plate-like co-crystals of purified recombinant N-terminally His-tagged TbTUT4 and UTP grow in 100 mM sodium cacodylate (pH 6.5), 200 mM calcium acetate, 18% (w/v) PEG-8000, crystal strucuture reveals two significantly different conformations of this TUTase:one molecule is in a relatively open apo confirmation, whereas the other displays a more compact TUTase-UTP complex	Trypanosoma brucei	Q381M1	675434
TUT4 in complex with UTP, X-ray diffraction structure analysis	Trypanosoma brucei	-	691064

pH STABILITY	pH STABILITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

TEMPERATURE STABILITY	TEMPERATURE STABILITY MAXIMUM	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
30	-	Vigna unguiculata	-	at least 3 h	643498

GENERAL STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
protein is stable at -20�C for several months	Leishmania tarentolae	-	675809

ORGANIC SOLVENT	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

OXIDATION STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

STORAGE STABILITY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
-70�C, 25% glycerol, stable	Homo sapiens	-	643500
4�C, does not tolerate more than several h at	Homo sapiens	-	643500
protein remains stable at -20�C in 50% glycerol up to 3 months	Trypanosoma brucei	-	675809

Purification/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
partial	Crithidia fasciculata	-	643501
2400fold to near homogeneity	Homo sapiens	-	643500
partial purification of U6 snRNA specific enzyme form via chromatography on a U6 snRNA affinity resin, separation from unspecific enzyme form	Homo sapiens	-	643504
homogenous RET1 is purified from an enriched mitochondrial fraction of Leishmania tarentolae by a series of chromatographic steps: affinity chromatography (polyU Sepharose 4B), anion-exchange chromatography (Poros 20 HQ 46/100 column), gel filtration (Superose 12 and Superose 6 columns) and hydrophobic interactions chromatography (Phenyl Superose column); recombinant RET1 from Leishmania tarentolae is purified by cation-exchange resin (Sepharose S Fast Flow), metal affinity chromatography (Talon metal affinity column), anion-exchange chromatography (Mono Q 5/5 column), expected yield: 0.2-0.5 mg/liter	Leishmania tarentolae	-	675809
expression and purification of 20S editosome-associated RET2 complex from Trypanosoma brucei by affinity isolation; recombinant RET1 from Trypanosoma brucei is purified by metal affinity chromatography (Talon metal affinity column), anion-exchange chromatography (HiTrap Q column) and loading onto a MonoS 5/5 column (final purification step), expected yield is 0.5 mg/liter of bacterial culture, it is important to maintain the concentration of KCl above 80 mM during purification, as protein tends to precipitate under low salt conditions; the recombinant expressed RET2 is purified by loading the protein extract onto a Talon affinity column and onto a HiTrap Sepharose S column, the expected protein yield is 1 mg of protein per liter of bacterial culture	Trypanosoma brucei	-	675809
recombinant enzyme	Trypanosoma brucei	-	661742
recombinant His-tagged enzyme from Escherichia coli	Trypanosoma brucei	-	643502, 643503
recombinant His-tagged wild-type and mutant TUT4 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ion exchange chromatography, and gel filtration	Trypanosoma brucei	Q381M1	675434
recombinant TAP-tagged enzyme from Leishmania tarentolae cells	Trypanosoma brucei	-	643505
614fold from uninfected and 117fold from CPMV-infected leaves	Vigna unguiculata	-	643498

Cloned/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
DNA sequence determination, gene is located on chromosome 9, expression as His-tagged protein in Escherichia coli	Leishmania tarentolae	-	643502
expression in Escherichia coli BL21(DE3) Codon Plus RIL, use of strains bearing additional tRNAs for expression of genes with G-rich codons is essential for RET1 expression	Leishmania tarentolae	-	675809
phylogenetic analysis, sequence comparison and signature motifs, overview	Leishmania tarentolae	-	691064
cloned in Trypanosoma brucei cells strain 29.13	Trypanosoma brucei	-	677097
DNA sequence determination, editosomal enzyme, expression as His-tagged protein in Escherichia coli	Trypanosoma brucei	-	643503
DNA sequence determination, mitochondrial enzyme, expression as His-tagged protein in Escherichia coli	Trypanosoma brucei	-	643502
expressed in Escherichia coli BL-21 as a 6xHis tagged fusion protein; TUT4, DNA and amino acid sequence determination and analysis, expression of N-terminally His-tagged wild-type and mutant TUT4 in Escherichia coli strain BL21(DE3)	Trypanosoma brucei	Q381M1	675434
expression in Escherichia coli BL21(DE3) Codon Plus RIL, use of strains bearing additional tRNAs for expression of genes with G-rich codons is essential for RET1 expression; for recombinant expression in Escherichia coli BL21 the RET2 gene lacking 21 codons at the 5' end, which correspond to the mitochondria importation signal, is cloned into pET15+vector to generate N-terminal 6 His-RET2 fusion protein; TbRET2 fused with the affinity TAP tag at the C-terminus using pLew79 vector in 29-13 cell line	Trypanosoma brucei	-	675809
expression of RET2 in Leishmania tarentolae cells as cytosolic enzyme by removing the mitochondrial targeting signal sequence	Trypanosoma brucei	-	643505
phylogenetic analysis, sequence comparison and signature motifs, overview	Trypanosoma brucei	-	691064

EXPRESSION	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

ENGINEERING	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
D136A	Trypanosoma brucei	Q381M1	inactive	675434
D297A	Trypanosoma brucei	Q381M1	Km=0.1728 (UTP), kcat=0.00033/sec (UTP)	675434
D297N	Trypanosoma brucei	Q381M1	Km=0.0139 (UTP), kcat=0.0000666/sec (UTP)	675434
D66A	Trypanosoma brucei	Q381M1	inactive	675434
D68A	Trypanosoma brucei	Q381M1	inactive	675434
E300A	Trypanosoma brucei	Q381M1	Km=0.3084 (UTP), kcat=0.0066/sec (UTP)	675434
N147A	Trypanosoma brucei	Q381M1	Km=0.1153 (UTP), kcat=0.005/sec (UTP)	675434
R121A	Trypanosoma brucei	Q381M1	Km=0.0028 (UTP), kcat=0.000033/sec (UTP),Km=0.132 (RNA), kcat= 0.005/sec (RNA)	675434
R121F	Trypanosoma brucei	Q381M1	inactive	675434
R126F	Trypanosoma brucei	Q381M1	inactive	675434
R141A	Trypanosoma brucei	Q381M1	Km=0.001 (UTP), kcat=0.00011/sec (UTP),Km=0.0726 (RNA), kcat= 0.0216/sec (RNA)	675434
R307A	Trypanosoma brucei	Q381M1	Km=0.0004 (UTP), kcat=0.000133/sec (UTP), Km=0.01 (RNA), kcat= 0.00833/sec (RNA)	675434
S148A	Trypanosoma brucei	Q381M1	Km=0.0873 (UTP), kcat=0.021/sec (UTP), Km=0.0002 (RNA), kcat= 0.0166/sec (RNA)	675434
S188A	Trypanosoma brucei	Q381M1	Km=0.034 (UTP), kcat=0.0005/sec (UTP), Km=0.0003 (RNA), kcat= 0.00166/sec (RNA)	675434
Y189A	Trypanosoma brucei	Q381M1	inactive	675434
Y189F	Trypanosoma brucei	Q381M1	Km=0.0213 (UTP), kcat=0.015/sec (UTP)	675434
F52A	Trypanosoma brucei	Q381M1	inactive	675434
additional information	Trypanosoma brucei	-	mutations of metal coordinating catalytic aspartic residues D66, D68 and D136, render TUT4 inactive but have no effect on UTP binding	691064

Renatured/COMMENTARY	ORGANISM	UNIPROT ACCESSION NO.	LITERATURE
No entries in this field

APPLICATION	ORGANISM	UNIPROT ACCESSION NO.	COMMENTARY	LITERATURE
No entries in this field

DISEASE	TITLE OF PUBLICATION	LINK TO PUBMED
No entries in this field

REF.	AUTHORS	TITLE	JOURNAL	VOL.	PAGES	YEAR	ORGANISM	LINK TO PUBMED	SOURCE
1997	Bakalara, N.; Simpson, A.M.; Simpson, L.	The Leishmania kinetoplast-mitochondrion contains terminal uridylyltransferase and RNA ligase activities	J. Biol. Chem.	264	18679-18686	1989	Leishmania tarentolae	PubMed
643498	Zabel, P.; Dorssers, L.; Wernars, K.; van Kammen, A.	Terminal uridylyl transferase of Vigna unguiculata: purification and characterization of an enzyme catalyzing the addition of a single UMP residue to the 3-end of an RNA primer	Nucleic Acids Res.	9	2433-2453	1981	Vigna unguiculata	PubMed
643500	Andrews, N.C.; Baltimore, D.	Purification of a terminal uridylyltransferase that acts as host factor in the in vitro poliovirus replicase reaction	Proc. Natl. Acad. Sci. USA	83	221-225	1986	Homo sapiens	PubMed
643501	Sugisaki, H.	Purification of terminal uridylyltransferase from the Crithidia kinetoplast-mitochondrion	ICR Annual Report	7	58-59	2001	Crithidia fasciculata	-
643502	Aphasizhev, R.; Sbicego, S.; Peris, M.; Jang, S.H.; Aphasizheva, I.; Simpson, A.M.; Rivlin, A.; Simpson, L.	Trypanosome mitochondrial 3' terminal uridylyl transferase (TUTase): the key enzyme in U-insertion/deletion RNA editing	Cell	108	637-648	2002	Leishmania tarentolae, Trypanosoma brucei	PubMed
643503	Ernst, N.L.; Panicucci, B.; Igo, R.P., Jr.; Panigrahi, A.K.; Salavati, R.; Stuart, K.	TbMP57 is a 3' terminal uridylyl transferase (TUTase) of the Trypanosoma brucei editosome	Mol. Cell	11	1525-1536	2003	Trypanosoma brucei	PubMed
643504	Trippe, R.; Richly, H.; Benecke, B.J.	Biochemical characterization of a U6 small nuclear RNA-specific terminal uridylyltransferase	Eur. J. Biochem.	270	971-980	2003	Homo sapiens	PubMed
643505	Aphasizhev, R.; Aphasizheva, I.; Simpson, L.	A tale of two TUTases	Proc. Natl. Acad. Sci. USA	100	10617-10622	2003	Trypanosoma brucei	PubMed
643506	Gott, J.M.	Two distinct roles for terminal uridylyl transferases in RNA editing	Proc. Natl. Acad. Sci. USA	100	10583-10584	2003	Trypanosoma brucei	PubMed
661742	Aphasizhev, R.; Aphasizheva, I.; Simpson, L.	Multiple terminal uridylyltransferases of trypanosomes	FEBS Lett.	572	15-18	2004	Trypanosoma brucei	PubMed
662323	Nebohacova, M.; Maslov, D.A.; Falick, A.M.; Simpson, L.	The effect of RNA interference down-regulation of RNA editing 3'-terminal uridylyl transferase (TUTase) 1 on mitochondrial de novo protein synthesis and stability of respiratory complexes in Trypanosoma brucei	J. Biol. Chem.	279	7819-7825	2004	Trypanosoma brucei	PubMed
675434	Stagno, J.; Aphasizheva, I.; Rosengarth, A.; Luecke, H.; Aphasizhev, R.	UTP-bound and Apo structures of a minimal RNA uridylyltransferase	J. Mol. Biol.	366	882-899	2007	Trypanosoma brucei	PubMed
675809	Aphasizhev, R.; Aphasizheva, I.	RNA editing uridylyltransferases of trypanosomatids	Methods Enzymol.	424	55-73	2007	Leishmania tarentolae, Trypanosoma brucei	PubMed
677097	Zhelonkina, A.G.; OHearn, S.F.; Law, J.A.; Cruz-Reyes, J.; Huang, C.E.; Alatortsev, V.S.; Sollner-Webb, B.	T. brucei RNA editing: action of the U-insertional TUTase within a U-deletion cycle	RNA	12	476-487	2006	Trypanosoma brucei	PubMed
691064	Aphasizhev, R.; Aphasizheva, I.	Terminal RNA uridylyltransferases of trypanosomes	Biochim. Biophys. Acta	1779	270-280	2008	Leishmania tarentolae, Trypanosoma brucei	PubMed

LINKS TO OTHER DATABASES (specific for EC-Number 2.7.7.52)

NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM

IUBMB Enzyme Nomenclature

PROSITE Database of protein families and domains

SYSTERS

Protein Mutant Database

InterPro (database of protein families, domains and functional sites)

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